Difference between revisions of "Team:NYU Shanghai/Protocols"
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</ul> | </ul> | ||
</p> | </p> | ||
+ | <p> | ||
<br>Lysis Buffer | <br>Lysis Buffer | ||
<p>For E. coli cell lysis, use a freshly prepared lysozyme solution (10 mg/ml) in 10 mM Tris-HCl, pH 8.0.</p> | <p>For E. coli cell lysis, use a freshly prepared lysozyme solution (10 mg/ml) in 10 mM Tris-HCl, pH 8.0.</p> | ||
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<br>Preparing 1mM D-Luciferin | <br>Preparing 1mM D-Luciferin | ||
<p>Directions for a 5 mg sample: Dissolve 0.034 gr dithiothreitol in 22 mL of QH2O. Add 0.2 mL of this DTT solution to the 5 mg d-luciferin. Add 4 uL of 10M NaOH to dissolve the luciferin. Dilute this into the remaining 21.8 mL of DTT solution and store as aliquots at - 80 ºC in darkness until use.</p> | <p>Directions for a 5 mg sample: Dissolve 0.034 gr dithiothreitol in 22 mL of QH2O. Add 0.2 mL of this DTT solution to the 5 mg d-luciferin. Add 4 uL of 10M NaOH to dissolve the luciferin. Dilute this into the remaining 21.8 mL of DTT solution and store as aliquots at - 80 ºC in darkness until use.</p> | ||
+ | </p> | ||
+ | <p> | ||
<br>Procedure<br> | <br>Procedure<br> | ||
Bacterial lysis: | Bacterial lysis: | ||
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<li>Light should be emitted within two-three seconds.</li> | <li>Light should be emitted within two-three seconds.</li> | ||
</ol> | </ol> | ||
+ | </p> | ||
+ | <p> | ||
<br>Example Calculations<br> | <br>Example Calculations<br> | ||
Lysis Buffer (Desired Total Volume: 15mL)<br> | Lysis Buffer (Desired Total Volume: 15mL)<br> | ||
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</tr> | </tr> | ||
</table> | </table> | ||
+ | </p> | ||
</p> | </p> | ||
</div> | </div> |
Revision as of 17:11, 16 September 2015